Point Mutations in EBV GH That Abrogate or Differentially Affect B Cell and Epithelial Cell Fusion

Overview

Journal Virology

Specialty Microbiology

Date 2007 Feb 20

PMID 17307213

Citations 30

Authors

Liguo Wu

Lindsey M Hutt-Fletcher

Affiliations

Soon will be listed here.

Abstract

Cell fusion mediated by Epstein-Barr virus requires three conserved glycoproteins, gB and gHgL, but activation is cell type specific. B cell fusion requires interaction between MHC class II and a fourth virus glycoprotein, gp42, which complexes non-covalently with gHgL. Epithelial cell fusion requires interaction between gHgL and a novel epithelial cell coreceptor and is blocked by excess gp42. We show here that gp42 interacts directly with gH and that point mutations in the region of gH recognized by an antibody that differentially inhibits epithelial and B cell fusion significantly impact both the core fusion machinery and cell-specific events. Substitution of alanine for glycine at residue 594 completely abrogates fusion with either B cells or epithelial cells. Substitution of alanine for glutamic acid at residue 595 reduces fusion with epithelial cells, greatly enhances fusion with B cells and allows low levels of B cell fusion even in the absence of gL.

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References

Moss B, Elroy-Stein O, Mizukami T, Alexander W, Fuerst T . Product review. New mammalian expression vectors. Nature. 1990; 348(6296):91-2. DOI: 10.1038/348091a0. View

Gianni T, Forghieri C, Campadelli-Fiume G . The herpesvirus glycoproteins B and H.L are sequentially recruited to the receptor-bound gD to effect membrane fusion at virus entry. Proc Natl Acad Sci U S A. 2006; 103(39):14572-7. PMC: 1600001. DOI: 10.1073/pnas.0606127103. View

Spear P, Longnecker R . Herpesvirus entry: an update. J Virol. 2003; 77(19):10179-85. PMC: 228481. DOI: 10.1128/jvi.77.19.10179-10185.2003. View

Borza C, Morgan A, Turk S, Hutt-Fletcher L . Use of gHgL for attachment of Epstein-Barr virus to epithelial cells compromises infection. J Virol. 2004; 78(10):5007-14. PMC: 400351. DOI: 10.1128/jvi.78.10.5007-5014.2004. View

Lopper M, Compton T . Coiled-coil domains in glycoproteins B and H are involved in human cytomegalovirus membrane fusion. J Virol. 2004; 78(15):8333-41. PMC: 446119. DOI: 10.1128/JVI.78.15.8333-8341.2004. View

Yaswen L, Stephens E, Davenport L, Hutt-Fletcher L . Epstein-Barr virus glycoprotein gp85 associates with the BKRF2 gene product and is incompletely processed as a recombinant protein. Virology. 1993; 195(2):387-96. DOI: 10.1006/viro.1993.1388. View

Whetter L, Day S, Brown E, Elroy-Stein O, Lemon S . Analysis of hepatitis A virus translation in a T7 polymerase-expressing cell line. Arch Virol Suppl. 1994; 9:291-8. DOI: 10.1007/978-3-7091-9326-6_29. View

Li Q, Turk S, Hutt-Fletcher L . The Epstein-Barr virus (EBV) BZLF2 gene product associates with the gH and gL homologs of EBV and carries an epitope critical to infection of B cells but not of epithelial cells. J Virol. 1995; 69(7):3987-94. PMC: 189130. DOI: 10.1128/JVI.69.7.3987-3994.1995. View

Galdiero M, Whiteley A, Bruun B, Bell S, Minson T, Browne H . Site-directed and linker insertion mutagenesis of herpes simplex virus type 1 glycoprotein H. J Virol. 1997; 71(3):2163-70. PMC: 191323. DOI: 10.1128/JVI.71.3.2163-2170.1997. View

10.

Li Q, Spriggs M, Kovats S, Turk S, Comeau M, Nepom B . Epstein-Barr virus uses HLA class II as a cofactor for infection of B lymphocytes. J Virol. 1997; 71(6):4657-62. PMC: 191687. DOI: 10.1128/JVI.71.6.4657-4662.1997. View

11.

Wang X, Hutt-Fletcher L . Epstein-Barr virus lacking glycoprotein gp42 can bind to B cells but is not able to infect. J Virol. 1998; 72(1):158-63. PMC: 109360. DOI: 10.1128/JVI.72.1.158-163.1998. View

12.

Wang X, Kenyon W, Li Q, Mullberg J, Hutt-Fletcher L . Epstein-Barr virus uses different complexes of glycoproteins gH and gL to infect B lymphocytes and epithelial cells. J Virol. 1998; 72(7):5552-8. PMC: 110204. DOI: 10.1128/JVI.72.7.5552-5558.1998. View

13.

Ferrer M, Kapoor T, Strassmaier T, Weissenhorn W, Skehel J, Oprian D . Selection of gp41-mediated HIV-1 cell entry inhibitors from biased combinatorial libraries of non-natural binding elements. Nat Struct Biol. 1999; 6(10):953-60. DOI: 10.1038/13324. View

14.

McShane M, Longnecker R . Cell-surface expression of a mutated Epstein-Barr virus glycoprotein B allows fusion independent of other viral proteins. Proc Natl Acad Sci U S A. 2004; 101(50):17474-9. PMC: 536015. DOI: 10.1073/pnas.0404535101. View

15.

Gianni T, Martelli P, Casadio R, Campadelli-Fiume G . The ectodomain of herpes simplex virus glycoprotein H contains a membrane alpha-helix with attributes of an internal fusion peptide, positionally conserved in the herpesviridae family. J Virol. 2005; 79(5):2931-40. PMC: 548475. DOI: 10.1128/JVI.79.5.2931-2940.2005. View

16.

Gianni T, Menotti L, Campadelli-Fiume G . A heptad repeat in herpes simplex virus 1 gH, located downstream of the alpha-helix with attributes of a fusion peptide, is critical for virus entry and fusion. J Virol. 2005; 79(11):7042-9. PMC: 1112143. DOI: 10.1128/JVI.79.11.7042-7049.2005. View

17.

Wu L, Borza C, Hutt-Fletcher L . Mutations of Epstein-Barr virus gH that are differentially able to support fusion with B cells or epithelial cells. J Virol. 2005; 79(17):10923-30. PMC: 1193614. DOI: 10.1128/JVI.79.17.10923-10930.2005. View

18.

Omerovic J, Lev L, Longnecker R . The amino terminus of Epstein-Barr virus glycoprotein gH is important for fusion with epithelial and B cells. J Virol. 2005; 79(19):12408-15. PMC: 1211543. DOI: 10.1128/JVI.79.19.12408-12415.2005. View

19.

Niwa H, Yamamura K, Miyazaki J . Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene. 1991; 108(2):193-9. DOI: 10.1016/0378-1119(91)90434-d. View